PDX1-expressing dorsal and ventral foregut endoderm

Patent metadata

Field	Value
Publication number	US-9499795-B2
Application number	US-201313962978-A
Country	US
Kind code	B2
Filing date	Aug 9, 2013
Priority date	Oct 27, 2005
Publication date	Nov 22, 2016
Grant date	Nov 22, 2016

Abstract

Disclosed herein are cell cultures comprising dorsal and/or ventral PDX1-positive foregut endoderm cells and methods of producing the same. Also disclosed herein are cell populations comprising substantially purified dorsal and/or ventral PDX1-positive foregut endoderm cells as well as methods for enriching, isolating and purifying dorsal and/or ventral PDX1-positive foregut endoderm cells from other cell types. Methods of identifying differentiation factors capable of promoting the differentiation of dorsal and/or ventral PDX1-positive foregut endoderm cells, are also disclosed.

First claim

What is claimed is:

1. An in vitro cell culture comprising human definitive endoderm cells, human pancreatic-duodenal homeobox factor-1 (PDX1) positive foregut endoderm cells, and a medium comprising an effective amount of a TGFβ superfamily member and an effective amount of a retinoid, wherein the effective amount of a TGFβ superfamily member and retinoid promote differentiation of definitive endoderm cells to PDX1-positive foregut endoderm cells, wherein the TGFβ superfamily member is activin A and wherein the retinoid is retinoic acid.

2. The cell culture of claim 1 , wherein at least a portion of the PDX1 positive foregut endoderm cells express the PDX1 protein.

3. The cell culture of claim 1 , wherein at least 10% of human cells in said in vitro culture are PDX1-positive foregut endoderm cells.

4. The cell culture of claim 1 , wherein at least 60% of human cells in said in vitro culture are PDX1-positive foregut endoderm cells.

5. The cell culture of claim 1 , wherein at least 75% of human cells in said in vitro culture are PDX1-positive foregut endoderm cells.

6. The cell culture of claim 1 , wherein the PDX1-positive foregut endoderm cells are selected from the group consisting of PDX1-positive dorsally-biased endoderm cells and PDX1-positive ventrally-biased endoderm cells.

7. The cell culture of claim 1 , wherein the PDX1-positive foregut endoderm cells are PDX1-positive dorsally-biased foregut endoderm cells.

8. The cell culture of claim 1 , wherein said PDX1-positive endoderm cells are derived in vitro from human pluripotent cells.

9. The cell culture of claim 8 , wherein said human pluripotent cells do not form embryoid bodies prior to differentiation.

10. The cell culture of claim 1 , wherein activin A is provided at a concentration of least 25 ng/ml.

11. The cell culture of claim 10 , wherein the retinoic acid is provided at a concentration of about 2 μM.

Assignees

• Viacyte Inc

Inventors

• D'Amour Kevin Allen
• Agulnick Alan D
• Eliazer Susan
• Baetge Emmanuel E

Classifications

• C12N2501/405

  Cell cycle regulated proteins, e.g. cyclins, cyclin-dependant kinases · CPC title

• C12N2501/16

  Activin; Inhibin; Mullerian inhibiting substance · CPC title

• C12N2501/41

  Hedgehog proteins; Cyclopamine (inhibitor) · CPC title

• C12N5/0606

  Pluripotent embryonic cells, e.g. embryonic stem cells [ES] (embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696) · CPC title

• C12N2501/155

  Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor · CPC title